Amphiphilic shuttle peptide delivers base editor ribonucleoprotein to correct the CFTR R553X mutation in well-differentiated airway epithelial cells.

Autor:	Kulhankova K; Department of Pediatrics, University of Iowa, Iowa City, IA, USA., Cheng AX; Feldan Therapeutics, Quebec, Qc, Canada., Traore S; Department of Pediatrics, University of Iowa, Iowa City, IA, USA., Auger M; Feldan Therapeutics, Quebec, Qc, Canada.; Department of Chemical Engineering, Laval University, Quebec, Qc, Canada., Pelletier M; Feldan Therapeutics, Quebec, Qc, Canada.; Department of Chemical Engineering, Laval University, Quebec, Qc, Canada., Hervault M; Feldan Therapeutics, Quebec, Qc, Canada., Wells KD; Division of Animal Sciences, Swine Somatic Cell Genome Editing Center, University of Missouri, Columbia, MO, USA., Green JA; Division of Animal Sciences, Swine Somatic Cell Genome Editing Center, University of Missouri, Columbia, MO, USA., Byrne A; Division of Animal Sciences, Swine Somatic Cell Genome Editing Center, University of Missouri, Columbia, MO, USA., Nelson B; Division of Animal Sciences, Swine Somatic Cell Genome Editing Center, University of Missouri, Columbia, MO, USA., Sponchiado M; Division of Animal Sciences, Swine Somatic Cell Genome Editing Center, University of Missouri, Columbia, MO, USA., Boosani C; Division of Animal Sciences, Swine Somatic Cell Genome Editing Center, University of Missouri, Columbia, MO, USA., Heffner CS; The Jackson Laboratory, Genetic Resource Science, Bar Harbor, ME, USA., Snow KJ; The Jackson Laboratory, Genetic Resource Science, Bar Harbor, ME, USA., Murray SA; The Jackson Laboratory, Genetic Resource Science, Bar Harbor, ME, USA., Villacreses RA; Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA, USA., Rector MV; Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA, USA.; Pappajohn Biomedical Institute, University of Iowa, Iowa City, IA, USA., Gansemer ND; Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA, USA.; Pappajohn Biomedical Institute, University of Iowa, Iowa City, IA, USA., Stoltz DA; Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA, USA.; Pappajohn Biomedical Institute, University of Iowa, Iowa City, IA, USA.; Department of Biomedical Engineering, University of Iowa, Iowa City, IA, USA.; Department of Molecular Physiology and Biophysics, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA, USA., Allamargot C; Central Microscopy Research Facility (CMRF), and Office for the Vice President of Research (OVPR), University of Iowa, Iowa City, IA, USA., Couture F; TransBIOTech, Lévis, Qc, Canada., Hemez C; Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of MIT and Harvard, Cambridge, MA, USA.; Graduate Program in Biophysics, Harvard University, Cambridge, MA, USA.; Howard Hughes Medical Institute, Harvard University, Cambridge, MA, USA., Hallée S; Feldan Therapeutics, Quebec, Qc, Canada., Barbeau X; Feldan Therapeutics, Quebec, Qc, Canada., Harvey M; Feldan Therapeutics, Quebec, Qc, Canada., Lauvaux C; Feldan Therapeutics, Quebec, Qc, Canada., Gaillet B; Department of Chemical Engineering, Laval University, Quebec, Qc, Canada., Newby GA; Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of MIT and Harvard, Cambridge, MA, USA.; Howard Hughes Medical Institute, Harvard University, Cambridge, MA, USA.; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.; Department of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, USA., Liu DR; Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of MIT and Harvard, Cambridge, MA, USA.; Howard Hughes Medical Institute, Harvard University, Cambridge, MA, USA.; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA., McCray PB Jr; Department of Pediatrics, University of Iowa, Iowa City, IA, USA., Guay D; Feldan Therapeutics, Quebec, Qc, Canada.; Department of Chemical Engineering, Laval University, Quebec, Qc, Canada.
Jazyk:	angličtina
Zdroj:	Nucleic acids research [Nucleic Acids Res] 2024 Oct 28; Vol. 52 (19), pp. 11911-11925.
DOI:	10.1093/nar/gkae819
Abstrakt:	Base editing could correct nonsense mutations that cause cystic fibrosis (CF), but clinical development is limited by the lack of delivery methods that efficiently breach the barriers presented by airway epithelia. Here, we present a novel amphiphilic shuttle peptide based on the previously reported S10 peptide that substantially improved base editor ribonucleoprotein (RNP) delivery. Studies of the S10 secondary structure revealed that the alpha-helix formed by the endosomal leakage domain (ELD), but not the cell penetrating peptide (CPP), was functionally important for delivery. By isolating and extending the ELD, we created a novel shuttle peptide, termed S237. While S237 achieved lower delivery of green fluorescent protein, it outperformed S10 at Cas9 RNP delivery to cultured human airway epithelial cells and to pig airway epithelia in vivo, possibly due to its lower net charge. In well-differentiated primary human airway epithelial cell cultures, S237 achieved a 4.6-fold increase in base editor RNP delivery, correcting up to 9.4% of the cystic fibrosis transmembrane conductance regulator (CFTR) R553X allele and restoring CFTR channel function close to non-CF levels. These findings deepen the understanding of peptide-mediated delivery and offer a translational approach for base editor RNP delivery for CF airway disease. (© The Author(s) 2024. Published by Oxford University Press on behalf of Nucleic Acids Research.)
Databáze:	MEDLINE
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